Protective Effects Of Zhen-Wu Tang On Expression Of PPAR? And ANGPTL4 Levels In IgA Nephropathy Rat Model

Background and ObjectivePrimary immunoglobulin A nephropathy(Primary Immunolobin A nephropthy,IgAN)is called IgA nephropathy,which is a group of glomerular mesangial with IgA deposition.IgA nephropathy accompanied by mesangial cell proliferation and mesangial matrix expansion as the main pathological changes of primary glomerulonephritis,the pathological mechanisms and therapeutic approach has been the focus of researchs.Peroxisome Prolifemtor-Activated Receptors gamma(PPAR y)is an important receptor involved in kidney disease occurs when its stimulated by drugs,it can affect the regulation of its downstream target genes,including angiopoietin-like protein 4(ANGPTL4),which is one of the downstream target genes of PPAR ?.ANGPTL4 is the latest discovery of a multifunctional secreted signaling protein factor which is closely associated with kidney disease.And TCM syndrome of IgA nephropathy is mainly spleen and kidney yang.Spleen deficiency shall impose yang and Water Treatment.Zhen-wu Tang(ZWT)is a traditional Chinese compound formula from Treatise on Febrile Diseases(Shanghan Lun)in the classic water yang compound.Previous experimental and clinical data proving it can be used in the treatment of IgAN,but its pharmacological mechanisms need to be further clarified.In this paper,we observe and study the protective effects of Zhen-wu Tang on IgAN rat model,so as to explore the therapeutic mechanism of Zhen-wu Tang on rat IgAN model whether or not related to the impact of regulatory role of PPAR? and ANGPTL4.In order to further verify the results of the experiment,this study experimental method used the drug blocked to better clarify the role in the treatment of IgAN links to explore and seek to develop new targets for prevention and treatment strategies IgAN.MethodsUsing bovine serum albumin(BSA)+ lipopolysaccharide(LPS)+ carbon tetrachloride(CCl4)with 12 weeks,and electing the stable IgAN experimental rats to treat with Zhen-wu Tang.1 Study the therapeutic effects of Zhen-wu Tang on IgAN rat model.Totally 60 SD male rats were randomly divided into six groups,namely the Control group,Model group,SYSWT group(0.864g/kg)and Zhen-wu Tang group with high(ZWT-H,16.8g/kg),medium(ZWT-M,8.4g/kg)and low(ZWT-L,4.2g/kg)with 10 rats in each group.Control group was not modeling process.Observe the general condition in the whole process.Coomassie Brilliant Blue and microscopy were used to examine 24h urinary protein excretion and the amount of urinary red blood cells,respectively.Levels of serum TP,ALB,BUN,SCr,SUA,TC,TG,HDL-C and LDL-C were evaluated.The prothrombin time(PT),activated partial thromboplastin time(APTT),thrombin time(TT),fiber Fibrinogen(FIB)were tested by blood coagulation analyzer.The platelet maximum aggregation rate(PAR),aggregative inhibition rate(AIR)and 5 min platelet depolymerization rate(PDR)were detected by Turbidimetric method.The glomerular deposition of IgA and C3 were measured by immunofluorescence.And the pathologic changes in renal tissue were observed by the hematoxylin and eosin staining.2 Investigate the mechanisms of PPAR? and ANGPTL4 in IgAN rats kidney with Zhen-wu Tang treatmentTotally 40 SD male rats were randomly divided into four groups,namely the Control group,Model group,Zhen-wu Tang group(ZWT,16.8g/kg),Zhen Wu Tang + GW9662(ZWT+GW9662,16.8g/kg+lmg/kg)with 10 rats of each group.Changes in mRNA expression of ANGPTL4 and PPAR y were measured by reverse transcriptase polymerase chain reaction,and protein expression of ANGPTL4 and PPAR? were measured by Western blotting.Results1 Therapeutic effects of Zhen-wu Tang on IgAN rat model.Compared with the model group,rats in ZWT-H,ZWT-M and ZWT-L groups possessed a better mental status,the weight,hair color,food intake and activity status were improved.The 24 h urinary protein excretion and urine red blood cell count in ZWT each groups were significantly lower(P<0.01).Rats in ZWT-H,ZWT-L groups the serum levels of TP were elevated(P<0.05).The serum ALB level in rats of ZWT-H group were significantly improved(P<0.05).Zhen-wu Tang each groups were no significant changes in BUN levels(P>0.05).The SCr levels decreased in ZWT-H rats(P<0.05).The SUA level in ZWT-H group was also decreased(P<0.01).Rats in ZWT each treatment group,the serum TC and serum TG levels were significantly decreased(P<0.01 or P<0.05).The level HDL-C in ZWT-H group was significantly increased(P<0.01),the level of LDL-C in ZWT each treatment group decreased significantly(P<0.01 or P<0.05).Rats in ZWT-H group,the levels of APTT,TT and FIB were significantly affected(P<0.01 or P<0.05),but they had no effect on the level of PT(P>0.05),and the therapeutic effect of ZWT-L group was not obvious.ZWT each groups of ADP-induced platelet aggregation have different degrees of inhibition(P<0.05),especially in the ZWT-H group,the most obvious inhibition rate of 10.67%,while an increase in platelet Zhen-wu Tang 5min depolymerization rate trend(P>0.05).Large ZWT-H group mouse serum IgA levels were significantly decreased(P<0.05),whereas serum C3 levels were significantly increased(P<0.05).Rats in each of ZWT treatment group shape and size has been restored,mesangial proliferative cells decreased matrix thickening and glomerular congestion eased most obvious effect of ZWT-H group.The glomerular IgA deposition in ZWT each treatment group were significantly reduced(P<0.01 or P<0.05),and the glomerular C3 deposition had a significant reduction in expression of each ZWT groups(P<0.01 or P<0.05).2 The mechanisms of PPAR? and ANCGPTL4 in IgAN rats kidney with Zhen-wu Tang treatment.PCR results showed that compared with the control group,expression of PPAR? mRNA in rats kidney of ZWT+GW9662 group was significantly inhibited(P<0.01),compared with the model and ZWT+GW9662 groups,the expression of PPAR? mRNA in ZWT group was significantly increased(P<0.01 or P<0.05).Compared with the control group,expression of ANGPTL4 mRNA in model and ZWT+GW9662 groups was significantly increased(P<0.01 or P<0.05),compared with the model and ZWT+GW9662 groups,expression of ANGP'TL4 mRNA in ZWT group significantly decreased(P<0.01 or P<0.05).Westernblot results showed that compared with the control group,the expression of PPAR?protein in rat kidney of ZWT+GW9662 significantly decreased(P<0.01),compared with the model and ZWT+ GW9662 groups,the expression of PPAR?protein in rat 'kidney of' ZWT group significantly increased(P<0.01).Compared with the control group,expression of ANGPTL4 protein in model and ZWT+GW9662 groups were significantly increased(P<0.01),compared with the model group and ZWT+GW9662 groups,the expression of ANGPTL4 protein in ZWT garoup was significantly decreased(P<0.01).Conclusions1 There is a significant protective effect of Zhen-wu Tang,which can significantly reduce proteinuria and hematuria,increased serum protein levels,lower serum lipid levels,reduce blood hypercoagulable state,improve renal function,and improve the renal damage,beside,a significant reduction in serum IgA level and improve complement C3 levels,reduce the deposition of IgA and C3 in the IgAN rat kidneys,thereby delaying the progression of the disease of IgAN.2 PPAR?/ANGPTL4 involved in the pathogenesis of IgAN.Zhen-wu Tang can significantly increase the expression of PPAR? IgAN rat kidneys,reduce expression of ANGPTL4,and improve serum lipid levels.These results suggest that ZWT possibly inhibits downstream target genes that ANGPTL4 expression through activation expression of PPAR? in the kidney was,and this molecular mechanisms of PPAR? and ANGPTL4 was achieve the therapeutic effect of IgAN rats.